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木薯贮藏根的大规模蛋白质组学及一个减少采后劣变的靶基因的鉴定

Large-Scale Proteomics of the Cassava Storage Root and Identification of a Target Gene to Reduce Postharvest Deterioration.

作者信息

Vanderschuren Hervé, Nyaboga Evans, Poon Jacquelyne S, Baerenfaller Katja, Grossmann Jonas, Hirsch-Hoffmann Matthias, Kirchgessner Norbert, Nanni Paolo, Gruissem Wilhelm

机构信息

Department of Biology, ETH Zurich, 8092 Zurich, Switzerland

Department of Biology, ETH Zurich, 8092 Zurich, Switzerland.

出版信息

Plant Cell. 2014 May;26(5):1913-1924. doi: 10.1105/tpc.114.123927. Epub 2014 May 29.

Abstract

Cassava (Manihot esculenta) is the most important root crop in the tropics, but rapid postharvest physiological deterioration (PPD) of the root is a major constraint to commercial cassava production. We established a reliable method for image-based PPD symptom quantification and used label-free quantitative proteomics to generate an extensive cassava root and PPD proteome. Over 2600 unique proteins were identified in the cassava root, and nearly 300 proteins showed significant abundance regulation during PPD. We identified protein abundance modulation in pathways associated with oxidative stress, phenylpropanoid biosynthesis (including scopoletin), the glutathione cycle, fatty acid α-oxidation, folate transformation, and the sulfate reduction II pathway. Increasing protein abundances and enzymatic activities of glutathione-associated enzymes, including glutathione reductases, glutaredoxins, and glutathione S-transferases, indicated a key role for ascorbate/glutathione cycles. Based on combined proteomics data, enzymatic activities, and lipid peroxidation assays, we identified glutathione peroxidase as a candidate for reducing PPD. Transgenic cassava overexpressing a cytosolic glutathione peroxidase in storage roots showed delayed PPD and reduced lipid peroxidation as well as decreased HO accumulation. Quantitative proteomics data from ethene and phenylpropanoid pathways indicate additional gene candidates to further delay PPD. Cassava root proteomics data are available at www.pep2pro.ethz.ch for easy access and comparison with other proteomics data.

摘要

木薯(Manihot esculenta)是热带地区最重要的块根作物,但收获后块根迅速发生的生理劣变(PPD)是木薯商业化生产的主要制约因素。我们建立了一种基于图像的PPD症状量化可靠方法,并使用无标记定量蛋白质组学来生成广泛的木薯块根和PPD蛋白质组。在木薯块根中鉴定出2600多种独特蛋白质,近300种蛋白质在PPD过程中表现出显著的丰度调节。我们鉴定了与氧化应激、苯丙烷生物合成(包括东莨菪素)、谷胱甘肽循环、脂肪酸α-氧化、叶酸转化和硫酸盐还原II途径相关的途径中的蛋白质丰度调节。谷胱甘肽相关酶(包括谷胱甘肽还原酶、谷氧还蛋白和谷胱甘肽S-转移酶)的蛋白质丰度和酶活性增加,表明抗坏血酸/谷胱甘肽循环起关键作用。基于蛋白质组学数据、酶活性和脂质过氧化测定的综合结果,我们鉴定谷胱甘肽过氧化物酶为减少PPD的候选物。在贮藏根中过表达胞质谷胱甘肽过氧化物酶的转基因木薯表现出PPD延迟、脂质过氧化减少以及HO积累降低。来自乙烯和苯丙烷途径的定量蛋白质组学数据表明还有其他基因候选物可进一步延迟PPD。木薯块根蛋白质组学数据可在www.pep2pro.ethz.ch上获取,便于访问并与其他蛋白质组学数据进行比较。

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